Palen T E, Cech T R
Nucleic Acids Res. 1983 Apr 11;11(7):2077-91. doi: 10.1093/nar/11.7.2077.
DNA renaturation kinetics was used to examine the relative accessibility of various regions of the Tetrahymena ribosomal RNA gene (rDNA) chromatin to micrococcal nuclease. In nuclei from cells active in rRNA transcription, the transcribed region of the rDNA chromatin was as much as 5-fold more accessible than the average of the total chromatin. As few as 20% inactive genes in the population could have accounted for all of the hybridization, so the transcribed region of the active units may be totally unprotected from nuclease degradation. The terminal non-transcribed spacer downstream from the transcription unit was also preferentially digested, but to a smaller degree. The central non-transcribed spacer was degraded to the same extent as total chromatin after a high degree of nuclease digestion. In nuclei from starved cells, which have 96% reduced rRNA transcription, the transcribed and terminal spacer regions of the rDNA were again more accessible than the total chromatin from the same nuclei, but the difference did not exceed 2-fold. We conclude that transcriptional activation is accompanied by major changes in the structure of the ribosomal gene chromatin, and that the extent and/or type of structural alteration differs in each functionally defined region of the rDNA.
利用DNA复性动力学来检测四膜虫核糖体RNA基因(rDNA)染色质的各个区域对微球菌核酸酶的相对可及性。在活跃进行rRNA转录的细胞的细胞核中,rDNA染色质的转录区域比总染色质的平均水平的可及性高多达5倍。群体中低至20%的非活性基因可能就解释了所有的杂交现象,因此活性单位的转录区域可能完全没有受到核酸酶降解的保护。转录单位下游的末端非转录间隔区也被优先消化,但程度较小。经过高度核酸酶消化后,中央非转录间隔区与总染色质的降解程度相同。在饥饿细胞的细胞核中,rRNA转录减少了96%,rDNA的转录和末端间隔区再次比来自相同细胞核的总染色质更易接近,但差异不超过2倍。我们得出结论,转录激活伴随着核糖体基因染色质结构的重大变化,并且在rDNA的每个功能定义区域中,结构改变的程度和/或类型有所不同。